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Fy 1991 Activities -. ^i i~~~~~~i:c:_ ~ ~ ~ ~ 99·B· 156:i;A ~~~~~~~~~i~~~~~~~~~~~~~~~~~j~~~~~~~~~~~~~~~~~W~~ ~ ~ ~ ~ ~~ ;~a~ r~r~a~i, ~_i: ?sd~,,ill~L~.O-s~:sl9 A* · ~~~~~~~~~~~l~~~~~j~~~~~L~~ *9 4~i ':A:i I~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ V ~~ ~ ~ ~ ~ ~ ~ ~ ~ UA 4 A ~ ~~ ~ ~~ ~ ~ ~~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~~~~~~~V'~~~~~~~~~~~~~~~~~~~~~ 4 A-A*4A 'AAV' 4 ~~~~~~~~~~~~~~~~ ,B This report has been reproduced directly from the best available copy. Available to DOE and DOE contractors from the Office of Scientific and Technical Informa- tion, P.O. Box 62, Oak Ridge, TN 37831; prices available from (615) 576-8401, FTS 626-8401. Available to the public from the National Technical Information Service, U.S. Department of Commerce, 5285 Port Royal Rd., Springfield, VA 22161. Cover Picture: The picture shows the filamentous cyanobacterium Anabaena PCC 7120 as seen by Nomarski optics following twenty-four hours of nitrogen deprivation. The mature nitrogen fixing heterocysts are seen as enlarged cells at regular intervals within the filaments. (Photo courtesy of Kurt Stepnitz) DOE/ER-0511P ANNUAL REPORT AND SUMMARIES OF FY 1991 ACTIVITIES DIVISION OF ENERGY BIOSCIENCES SEPTEMBER 1991 U.S. DEPARTMENT OF ENERGY OFFICE OF ENERGY RESEARCH OFFICE OF BASIC ENERGY SCIENCES DIVISION OF ENERGY BIOSCIENCES WASHINGTON, D.C. 20585 Program Overview of the Division of Energy Biosciences At a time when the issues of energy resources for the future, environmental quality, and U.S. competitiveness in high technology markets are all high on the list of national priorities, research that is oriented towards contributing to the long-term solutions of these concerns is generally recognized to occupy a significant role. This is one of the conclusions of the National Energy Strategy document released in 1990. As a component of the Department of Energy, the Energy Biosciences (EB) program of the Office of Basic Energy Sciences supports long-term research aimed at addressing energy-related problems utilizing biological systems. The EB program generates critical fundamental biological data, principally about plants and microorganisms, that contribute toward establishing the bases for future biotechnologies related to energy matters. The studies supported by EB are broad- ranging, yet focus on understanding basic principles and mechanisms rather than on the optimization of processes, which is in the realm of other Federal programs and industry. However, one important facet of the program is to encourage interactions with, and transfer of information to, commercial interests and more applied research programs. The ability of biological systems to perform myriads of chemical transformations resulting in constituents with unique properties has become progressively more evident in recent years. Over history, humankind has continued to exploit and modify biological systems for advantage. In recent years, enormous frontiers have been opened for utilization of biological processes as "biotechnologies" through the acquisition of crucial fundamental knowledge about biological systems. This has been facilitated particularly by discoveries on manipulating biosystems at the genetic level in ways previously unimagined. The primary challenge is to use the current and forthcoming information in a creative and prudent fashion in devising new technologies that will serve the needs of society without imposing undue risks to the health of its people and the natural environment which has suffered so seriously from some of man's other endeavors. The EB program is oriented towards important research areas that are, in some cases, underpopulated and not well defined in either an intellectual framework or in potential approaches to the problem area. The program seeks projects that attempt to break new ground in providing new insights into the basic mechanisms of how plants and non- medical microorganisms grow, survive, metabolize, differentiate, inherit traits and reproduce. This type of knowledge is critically important to fully integrate biology into the building of future technologies (biotechnology) aimed at serving society in an energy context. - i - The EB program content is broad in scope and may be broken down into the three categories cited below. Despite the divisions noted below, it is clear that the topics must and do integrate from the standpoint of natural processes, as well as in the formulation of approaches and insights in the research supported. The components of the EB program consist of: I. Primary Biological Energy Conversion. This category comprises research on plant and microbial photosynthesis, the solar energy driven process central to the support of life on earth. This category includes studies on the initial carbon dioxide fixing mechanisms as well as the associated water splitting and other component reactions. The program also supports research on the fundamental processes, such as the control of growth and development, that ultimately govern the form and amount of biomass a plant produces. In addition, investigations on plant interactions with the natural environment (including stress reactions, as well as the interactions with biological agents such as pathogens) are included in this category. Constituents of this broad research category include biophysical, biochemical and physiological, and genetic investigations. II. Bioconversion of Products. The utilization of the products of the primary energy conversion process is the nature of this second category. A prime component is studies on the diverse metabolic capabilities of plants and microorganisms in synthesizing materials that may ultimately be utilized for fuels and chemicals. These fundamental investigations focus on understanding pathways of metabolism and the genetic and biochemical regulatory mechanisms that determine the nature and amount of metabolic compounds converted. Of special interest are the synthesis, structure and function of plant cell walls, the predominant renewable biomass resource. Also of interest are the mechanisms by which lignocellulose of plant cell walls is degraded biologically into compounds of potential utility. Associated with this category is how plant-microbial interactions occur in symbiotic energy exchanges. A substantial amount of this category covers fundamental microbial conversions, such as methanogenesis and other diverse fermentative pathways. III. Infrastructure. Underpinning of the development of future biotechnologies is a key objective of the EB program. In this category are efforts to better understand genetic mechanisms involving the transfer of genetic information and its expression in plant and microbial species that oftentimes have not been studied very extensively. Also under study are genetic mechanisms in better known species such as maize where certain phenomena were first discovered. Further, in this portion of the program are such activities as the development of critical data bases, specific techniques and instrumentation. A training component for nurturing areas that are important but underpopulated is also encompassed. - ii- Representative of some of the activities sponsored by EB are the following: Mutant Analysis of Photosystem I of Photosynthesis If a process is to be analyzed genetically, by use of mutants in which that process is altered or defective, it often helps to find conditions in which the process is not needed. The photosynthetic activity of plants and cyanobacteria (once called blue-green algae) is dependent upon the activity of two light-utilizing systems, known as photosystem I and photosystem II. Using recombinant DNA techniques and the genetically manipulable unicellular cyanobacterium, Synechocystis, which can be grown without photosystem II, investigators succeeded several years ago in clarifying the nature of the electron donor to photosystem II. However, because the activity of photosystem I is essential for the growth of every photosynthetic organism that could be easily manipulated genetically, genetic analysis of photosystem I had progressed very little. That situation has now changed with the progress made by two groups at the Michigan State University Plant Research Laboratory (PRL). It has been shown that Synechocystis may be grown without photosynthesis if it be given only five minutes of photosynthetically inactive blue light per day; the group was thereupon able to mutagenize a polypeptide at the very core of photosystem I. Another group, in collaboration with researchers from Germany and Spain, showed that a strain of the filamentous cyanobacterium, Anabaena, that can grow in complete darkness, can also be mutated in chosen genes by the use of recombinant DNA techniques and transfer of altered DNA from Escherichia coli to Anabaena. These findings are enabling researchers in the PRL and elsewhere to define photosystem I with a more detailed analysis and thus enhance the overall understanding of the structure and function of the photosynthetic apparatus. Mechanisms of Plant/Fungal Pathogen Interactions Fungal toxins that are highly specific to a particular host plant have proven agriculturally devastating, for example, in the Southern Corn Leaf Blight epidemic of 1970. Although several such toxins were known, the biosynthetic apparatus responsible for their synthesis has been elusive. Moreover, in no instance was it precisely known why closely related strains of the same host plant were resistantto the fungal toxin. Both of these questions have recently been broached mainly through the efforts of researchers at the PRL. First,
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